The TGB1 Movement Protein of<i>Potato virus X</i>Reorganizes Actin and Endomembranes into the X-Body, a Viral Replication Factory

Tilsner, Jens; Linnik, Olga; Wright, Kathryn M.; Bell, Karen; Roberts, Alison G.; Lacomme, Christophe; Cruz, Simon Santa; Oparka, Karl J.

doi:10.1104/pp.111.189605

Cited by 119 publications

(180 citation statements)

References 74 publications

(155 reference statements)

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“…Pumilio-based RNA imaging has enabled subcellular detection of the mRNAs of mitochondrial NADH dehydrogenase and -actin, as well as retroviral RNA in mammalian cultured cells (20)(21)(22)(23), and of the genomes of Tobacco mosaic virus, Potato virus X and Turnip mosaic virus in live plant tissue (24)(25)(26)(27). Of particular relevance to the topic of this volume, Pumilio-based RNA imaging has recently shown that Potato virus X is present in membrane structures at the entrances of plasmodesmata that also contain the viral replicase (27).…”

Section: Rna-binding Domain Of Pumilio Proteinsmentioning

confidence: 99%

“…This principle is the only form of Pumilio-based RNA imaging so far applied in plants ( [24][25][26][27].…”

Section: Choice Of Reporter Systemmentioning

confidence: 99%

“…PUM-BiFC imaging can be used in two different modes: PUMHDs can be modified to bind to the native RNA of interest (20,21,(23)(24)(25), or the RNA can be tagged with recognition motifs of previously characterised PUMHD variants (22,24,26,27,33) (Table 1). As described in the introduction, leaving the RNA unmodified has the advantage of avoiding potential RNA processing and localisation artefacts that could be caused by a tag, and by the expression of the tagged construct from an artificial genomic context.…”

Section: Choice Of Target Sequences and Pumhd Variantsmentioning

confidence: 99%

“…In principle, PUM-BiFC reporter constructs can be stably expressed, but so far all Pumilio-based RNA imaging in plants has been done with transient expression systems (agroinfiltration or microprojectile bombardment) (24)(25)(26)(27). Since nonspecific background fluorescence increases with the accumulation of the PUM-BiFC fusions, transient expression may be preferable as it permits limiting background fluorescence by reducing the expression time (see 3.3.…”

Section: Choice Of Expression Systemmentioning

confidence: 99%

“…After these general considerations, the following protocol describes the construction of a PUMBiFC system to image the untagged, genomic RNA of Tobacco mosaic virus (TMV) (24,26 Table 2). …”

Section: Choice Of Expression Systemmentioning

confidence: 99%

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Pumilio-Based RNA In Vivo Imaging

Tilsner

2014

Methods in Molecular Biology

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SummarySubcellular, sequence-specific detection of RNA in vivo is a powerful tool to study the macromolecular transport that occurs through plasmodesmata. The RNA binding domain of Pumilio proteins can be engineered to bind RNA sequences of choice and fused to fluorescent proteins for RNA imaging. This chapter describes the construction of a Pumilio-based imaging system to track the RNA of Tobacco mosaic virus in vivo, and practical aspects of RNA live-cell imaging.

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Section: Rna-binding Domain Of Pumilio Proteinsmentioning

confidence: 99%

“…This principle is the only form of Pumilio-based RNA imaging so far applied in plants ( [24][25][26][27].…”

Section: Choice Of Reporter Systemmentioning

confidence: 99%

Section: Choice Of Target Sequences and Pumhd Variantsmentioning

confidence: 99%

Section: Choice Of Expression Systemmentioning

confidence: 99%

Section: Choice Of Expression Systemmentioning

confidence: 99%

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Pumilio-Based RNA In Vivo Imaging

Tilsner

2014

Methods in Molecular Biology

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Potexviruses and Carlaviruses – Short Filamentous Viruses

Rubino

2022

Encyclopedia of Life Sciences

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Potexviruses and carlaviruses are plant‐infecting viruses belonging to the family Aphaflexiviridae and Betaflexiviridae , respectively. Both have slightly flexuous filamentous particles measuring 470–580 × 13 nm (potexviruses), and 610–700 × 12–15 nm (carlaviruses). Virions contain 94–95% protein and 5–6% positive sense, single‐stranded ribonucleic acid c . 5800–7000 nucleotides (nt) (potexviruses) and 7900–9000 nt (carlaviruses) in size. Both virus types have a genome with a similar structural organisation, comprising five (potexviruses) or six (carlaviruses) open reading frames, three of which, known as the ‘triple gene block’, encode polypeptides that facilitate virus movement within the host. Natural hosts of both virus types are herbaceous (weeds, ornamentals, vegetables) rather than woody plants. Many potexviruses are agents of severe diseases, whereas a high percentage of carlaviruses induce symptomless infections. Transmission of potexviruses in nature is, by and large, by mechanical contact, whereas carlaviruses are transmitted primarily by aphids with a nonpersistent modality. Key Concepts Potexviruses and carlaviruses infect primarily herbaceous hosts after mechanical or vector‐driven transmission, respectively. Particles of both genera are filamentous and slightly flexuous and accumulate in aggregates in the cytoplasm of parenchyma cells. Demarcation of species within the genera takes into account molecular and biological criteria. Both genera are characterised by the presence of the so‐called triple gene block that provides the support for virus movement in infected hosts. Potexviruses and carlaviruses constitute models for virus replication, movement and encapsidation studies. Control is based on preventive measures, such as sanitary selection, production of virus‐free seed lots and resistance through conventional breeding and genetic engineering.

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The potential of engineered eukaryotic RNA‐binding proteins as molecular tools and therapeutics

2019

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Eukaroytic RNA‐binding proteins (RBPs) recognize and process RNAs through recognition of their sequence motifs via RNA‐binding domains (RBDs). RBPs usually consist of one or more RBDs and can include additional functional domains that modify or cleave RNA. Engineered RBPs have been used to answer basic biology questions, control gene expression, locate viral RNA in vivo, as well as many other tasks. Given the growing number of diseases associated with RNA and RBPs, engineered RBPs also have the potential to serve as therapeutics. This review provides an in depth description of recent advances in engineered RBPs and discusses opportunities and challenges in the field. This article is categorized under: RNA Interactions with Proteins and Other Molecules > Protein–RNA Recognition RNA Methods > RNA Nanotechnology RNA in Disease and Development > RNA in Disease

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The TGB1 Movement Protein ofPotato virus XReorganizes Actin and Endomembranes into the X-Body, a Viral Replication Factory

Cited by 119 publications

References 74 publications

Pumilio-Based RNA In Vivo Imaging

Pumilio-Based RNA In Vivo Imaging

Potexviruses and Carlaviruses – Short Filamentous Viruses

The potential of engineered eukaryotic RNA‐binding proteins as molecular tools and therapeutics

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